CCR-3 receptor antagonists (I)

ABSTRACT

The invention provides compounds of Formula (I):  
                 
 
     wherein: R 1 -R 4 , A, D, and L have any of the values defined in the specification that are CCR-3 receptor antagonists, pharmaceutical compositions containing them, methods for their use, and methods and intermediates useful for preparing them.

[0001] This application incorporates by reference U.S. Provisionalapplication Serial No. 60/334,653 “Piperidine CCR-3 ReceptorAntagonists” Attorney Docket R0123A-PRO, and U.S. Provisionalapplication Serial No. 60/334,655 “Piperazine CCR-3 ReceptorAntagonists” Attorney Docket R0124A-PRO, both filed concurrently on Nov.30, 2001; issued U.S. Pat. No. 6,323,223, and issued U.S. Pat. No.6,166,015. This application also claims the benefit of priority of U.S.Provisional Patent Application Serial No. 60/334,819, filed Nov. 30,2001, which is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

[0002] The invention relates to CCR-3 receptor antagonists,pharmaceutical compositions containing them, methods for their use, andmethods and intermediates useful for preparing them.

BACKGROUND INFORMATION

[0003] Tissue eosinophilia is a feature of a number of pathologicalconditions such as asthma, rhinitis, eczema and parasitic infections(see Bousquet, J. et al., N. Eng. J. Med. 323: 1033-1039 (1990) and Kay,A. B. and Corrigan, C. J., Br. Med. Bull. 48:51-64 (1992)). In asthma,eosinophil accumulation and activation are associated with damage tobronchial epithelium and hyperresponsiveness to constrictor mediators.Chemokines such as RANTES, eotaxin and MCP-3 are known to activateeosinophils (see Baggiolini, M. and Dahinden, C. A., Immunol. Today.15:127-133 (1994), Rot, A. M. et al., J. Exp. Med. 176, 1489-1495 (1992)and Ponath, P. D. et al., J. Clin. Invest., Vol. 97, #3, 604-612(1996)). However, unlike RANTES and MCP-3 which also induce themigration of other leukocyte cell types, eotaxin is selectivelychemotactic for eosinophils (see Griffith-Johnson, D. A. et al.,Biochem. Biophy. Res. Commun. 197:1167 (1993) and Jose, P. J. et al.,Biochem. Biophy. Res. Commun. 207, 788 (1994)). Specific eosinophilaccumulation was observed at the site of administration of eotaxinwhether by intradermal or intraperitoneal injection or aerosolinhalation (see Griffith-Johnson, D. A. et al., Biochem. Biophy. Res.Commun. 197:1167 (1993); Jose, P. J. et al., J. Exp. Med. 179, 881-887(1994); Rothenberg, M. E. et al., J. Exp. Med. 181, 1211 (1995) andPonath, P. D., J. Clin. Invest., Vol. 97, #3, 604-612 (1996)).

[0004] Glucocorticoids such as dexamethasone, methprednisolone andhydrocortisone have been used for treating many eosinophil-relateddisorders, including bronchial asthma (R. P. Schleimer et al., Am. Rev.Respir. Dis., 141, 559 (1990)). The glucocorticoids are believed toinhibit IL-5 and IL-3 mediated eosinophil survival in these diseases.However, prolonged use of glucocorticoids can lead to side effects suchas glaucoma, osteoporosis and growth retardation in the patients (seeHanania, N. A. et al., J. Allergy and Clin. Immunol., Vol. 96, 571-579(1995) and Saha, M. T. et al., Acta Paediatrica, Vol. 86, #2, 138-142(1997)). It is therefore desirable to have an alternative means oftreating eosinophil related diseases without incurring these undesirableside effects.

[0005] Recently, the CCR-3 receptor was identified as a major chemokinereceptor that eosinophils use for their response to eotaxin, RANTES andMCP-3. When transfected into a murine pre-beta. lymphoma line, CCR-3bound eotaxin, RANTES and MCP-3 conferred chemotactic responses on thesecells to eotaxin, RANTES and MCP-3 (see Ponath, P. D. et al., J. Exp.Med. 183, 2437-2448 (1996)). The CCR-3 receptor is expressed on thesurface of eosinophils, T-cells (subtype Th-2), basophils and mast cellsand is highly selective for eotaxin. Studies have shown thatpretreatment of eosinophils with an anti-CCR-3 mAb completely inhibitseosinophil chemotaxis to eotaxin, RANTES and MCP-3 (see Heath, H. etal., J. Clin. Invest., Vol. 99, #2, 178-184 (1997)). Applicants' issuedU.S. Pat. Nos. 6,140,344 and 6,166,015 and published EP applicationEP903349, published Mar. 24, 1999 disclose CCR-3 antagonists thatinhibit eosinophilic recruitment by chemokine such as eotaxin.

[0006] Therefore, blocking the ability of the CCR-3 receptor to bindRANTES, MCP-3 and eotaxin and thereby preventing the recruitment ofeosinophils should provide for the treatment of eosinophil-mediatedinflammatory diseases.

SUMMARY OF THE INVENTION

[0007] The present invention concerns compounds which are capable ofinhibiting the binding of eotaxin to the CCR-3 receptor and therebyprovide a means of combating eosinophil induced diseases, such asasthma.

[0008] In a first aspect, this invention provides a compound of Formula(I):

[0009] wherein:

[0010] R¹ is methylene or ethylene;

[0011] R² is optionally substituted phenyl;

[0012] R³ is hydrogen, alkyl, acyl, aryl, or arylalkyl;

[0013] ring A is a cycloalkyl, heterocyclyl, or optionally substitutedphenyl;

[0014] D is N or C—R^(b);

[0015] L is —C(═O)—, —C(═S)—, —SO₂—, —C(═O)N(R_(a))—, —C(═S) N(R_(a))—,—SO₂ N(R_(a))—, —C(═O)O—, —C(═S)O—, —S(═O)₂O—;

[0016] R⁴ is alkyl, cycloalkyl, alkenyl, alkynyl, heteroalkyl oracylalkyl;

[0017] R_(a) is hydrogen, alkyl, acyl, aryl, arylalkyl, alkoxycarbonyl,or benzyloxycarbonyl; and

[0018] R^(b) is hydrogen or alkyl;

[0019] and prodrugs, individual isomers, racemic and non-racemicmixtures of isomers, and pharmaceutically acceptable salts and solvatesthereof.

[0020] In a second aspect, this invention provides pharmaceuticalcompositions containing a therapeutically effective amount of a compoundof Formula (I) or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable excipient.

[0021] In a third aspect, this invention provides a method of treatmentof a disease in a mammal treatable by administration of a CCR-3 receptorantagonist, comprising administration of a therapeutically effectiveamount of a compound of Formula (I) or a pharmaceutically acceptablesalt thereof. The disease states include respiratory diseases such asasthma.

[0022] In a fourth aspect, this invention provides processes disclosedherein for preparing compounds of Formula (I).

[0023] In a fifth aspect, this invention provides novel intermediatesdisclosed herein that are useful for preparing compounds of Formula (I).

[0024] In a sixth aspect, this invention provides a compound of Formula(I) or a pharmaceutically acceptable salt thereof for use in medicaltherapy or diagnosis (e.g. for treating asthma).

[0025] In a seventh aspect, this invention provides the use of acompound of Formula (I) or a pharmaceutically acceptable salt thereoffor the manufacture of a medicament useful for treating a disease in amammal treatable by administration of a CCR-3 receptor antagonist (e.g.asthma).

DETAILED DESCRIPTION OF THE INVENTION

[0026] Definitions

[0027] Unless otherwise stated, the following terms used in thespecification and claims have the meanings given below.

[0028] “Acyl” means a radical —C(O)R, where R is hydrogen, alkyl,cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl wherein alkyl,cycloalkyl, cycloalkylalkyl, and phenylalkyl are as defined herein.Representative examples include, but are not limited to formyl, acetyl,cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl,and the like.

[0029] “Acylalkyl” means a radical -alkylene-C(O)R where R is hydrogen,alkyl, haloalkyl, cycloalkyl, cycloalkyl-alkyl, optionally substitutedphenyl, benzyl, hydroxy, alkoxy, amino, monoalkylamino or dialkylamino.Representative examples include methylcarbonyl-methyl,2-(ethoxycarbonyl)ethyl, 2-(methoxycarbonyl)ethyl, 2-carboxyethyl andthe like.

[0030] “Acylamino” means a radical —NR′C(O)R, where R′ is hydrogen oralkyl, and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl orphenylalkyl wherein alkyl, cycloalkyl, cycloalkylalkyl, and phenylalkylare as defined herein. Representative examples include, but are notlimited to formylamino, acetylamino, cylcohexylcarbonylamino,cyclohexylmethylcarbonylamino, benzoylamino, benzylcarbonylamino, andthe like.

[0031] “Alkoxy” means a radical —OR where R is an alkyl as definedherein e.g., methoxy, ethoxy, propoxy, butoxy and the like.

[0032] “Alkoxycarbonyl” means a radical —C(O)—R where R is alkoxy is asdefined herein.

[0033] “Alkenyl” means a linear monovalent hydrocarbon radical of two tosix carbon atoms or a branched monovalent hydrocarbon radical of threeto six carbon atoms, containing at least one double bond, e.g., ethenyl,propenyl, and the like.

[0034] “Alkyl” means a linear saturated monovalent hydrocarbon radicalof one to six carbon atoms or a branched saturated monovalenthydrocarbon radical of three to six carbon atoms, e.g., methyl, ethyl,propyl, 2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like.

[0035] “Alkylamino” or “Monoalkylamino” means a radical —NHR where Rrepresents an alkyl, cycloalkyl or cycloalkyl-alkyl group as definedherein. Representative examples include, but are not limited tomethylamino, ethylamino, isopropylamino, cyclohexylamino, and the like.

[0036] “Alkylene” means a linear saturated divalent hydrocarbon radicalof one to six carbon atoms or a branched saturated divalent hydrocarbonradical of three to six carbon atoms, e.g., methylene, ethylene,2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene,and the like.

[0037] “Alkynyl” means a linear monovalent hydrocarbon radical of two tosix carbon atoms or a branched monovalent hydrocarbon radical of threeto six carbon atoms, containing at least one triple bond, e.g., ethynyl,propynyl, and the like.

[0038] “Alkylsulfonyl” means a radical —S(O)₂R where R is an alkyl,cycloalkyl or cycloalkyl-alkyl group as defined herein, e.g.,methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl,cyclohexylsulfonyl and the like.

[0039] “Alkylsulfinyl” means a radical —S(O)R where R is an alkyl,cycloalkyl or cycloalkyl-alkyl group as defined herein e.g.,methylsulfinyl, ethylsulfinyl, propylsulfinyl, butylsulfinyl,cyclohexylsulfinyl and the like.

[0040] “Alkylthio” means a radical —SR where R is an alkyl as definedabove e.g., methylthio, ethylthio, propylthio, butylthio, and the like.

[0041] “Aryl” means a monocyclic or bicyclic aromatic hydrocarbonradical which is optionally substituted with one or more substituents,preferably one, two or three, substituents preferably selected from thegroup consisting of alkyl, haloalkyl, hydroxyalkyl, heteroalkyl, acyl,acylamino, amino, alkylamino, dialkylamino, alkylthio, alkylsulfinyl,alkylsulfonyl, —SO₂NR′R″ (where R′ and R″ are independently hydrogen oralkyl), alkoxy, haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halo,nitro, cyano, mercapto, methylenedioxy or ethylenedioxy. Morespecifically the term aryl includes, but is not limited to, phenyl,chlorophenyl, fluorophenyl, methoxyphenyl, 1-naphthyl, 2-naphthyl, andthe derivatives thereof.

[0042] “Arylene” means a divalent aryl group as defined above.

[0043] “Arylalkyl” refers to an alkyl radical as defined herein in whichone of the hydrogen atoms of the alkyl group is replaced with an arylgroup. Typical arylalkyl groups include, but are not limited to, benzyl,2-phenylethan-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like.

[0044] “Aryloxy” means a radical —O—R where R is an aryl group asdefined herein.

[0045] “Carbamoyl” means the radical —C(═O)NH₂.

[0046] “Cycloalkyl” refers to a saturated monovalent cyclic hydrocarbonradical of three to seven ring carbons e.g., cyclopropyl, cyclobutyl,cyclohexyl, 4-methylcyclohexyl, and the like.

[0047] “Cycloalkyl-alkyl” means a radical —R^(x)R^(y) where R^(x) is analkylene group and R^(y) is cycloalkyl group as defined herein, e.g.,cyclohexylmethyl, and the like.

[0048] “Dialkylamino” means a radical —NRR′ where R and R′ independentlyrepresent an alkyl, cycloalkyl, or cycloalkylalkyl group as definedherein. Representative examples include, but are not limited todimethylamino, methylethylamino, di(1-methylethyl)amino,(cyclohexyl)(methyl)amino, (cyclohexyl)(ethyl)amino,(cyclohexyl)(propyl)amino, (cyclohexylmethyl)(methyl)amino,(cyclohexylmethyl)(ethyl)amino, and the like.

[0049] “Halo” means fluoro, chloro, bromo, or iodo, preferably fluoroand chloro.

[0050] “Haloalkyl” means alkyl substituted with one or more same ordifferent halo atoms, e.g., —CH₂Cl, —CF₃, —CH₂CF₃, —CH₂CCl₃, and thelike.

[0051] “Heteroaryl” means a monocyclic or bicyclic radical of 5 to 12ring atoms having at least one aromatic ring containing one, two, orthree ring heteroatoms selected from N, O, or S, the remaining ringatoms being C, with the understanding that the attachment point of theheteroaryl radical will be on an aromatic ring. The heteroaryl ring isoptionally substituted independently with one or more substituents,preferably one or two substituents, selected from alkyl, haloalkyl,hydroxyalkyl, heteroalkyl, acyl, acylamino, amino, alkylamino,dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl, —SO₂NR′R″ (whereR′ and R″ are independently hydrogen or alkyl), alkoxy, haloalkoxy,alkoxycarbonyl, carbamoyl, hydroxy, halo, nitro, cyano, mercapto,methylenedioxy or ethylenedioxy. More specifically the term heteroarylincludes, but is not limited to, pyridyl, furanyl, thienyl, thiazolyl,isothiazolyl, triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl,pyrimidinyl, benzofuranyl, tetrahydrobenzofuranyl, isobenzofuranyl,benzothiazolyl, benzoisothiazolyl, benzotriazolyl, indolyl, isoindolyl,benzoxazolyl, quinolyl, tetrahydroquinolinyl, isoquinolyl,benzimidazolyl, benzisoxazolyl or benzothienyl and derivatives thereof.

[0052] “Heteroarylene” means a divalent heteroaryl group as definedabove.

[0053] “Heteroarylalkyl means an alkyl radical as defined herein inwhich one of the hydrogen atoms of the alkyl group is replaced with aheteroaryl group.

[0054] “Heteroalkyl” means an alkyl radical as defined herein whereinone, two or three hydrogen atoms have been replaced with a substituentindependently selected from the group consisting of —OR^(a),—NR^(b)R^(c), and —S(O)_(n)R^(d) (where n is an integer from 0 to 2),with the understanding that the point of attachment of the heteroalkylradical is through a carbon atom, wherein R^(a) is hydrogen, acyl,alkyl, cycloalkyl, or cycloalkylalkyl; R^(b) and R^(c) are independentlyof each other hydrogen, acyl, alkyl, cycloalkyl, or cycloalkylalkyl;when n is 0, R^(d) is hydrogen, alkyl, cycloalkyl, or cycloalkylalkyl,and when n is 1 or 2, R^(d) is alkyl, cycloalkyl, cycloalkylalkyl,amino, acylamino, monoalkylamino, or dialkylamino. Representativeexamples include, but are not limited to, 2-hydroxyethyl,3-hydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxypropyl,1-hydroxymethylethyl, 3-hydroxybutyl, 2,3-dihydroxybutyl,2-hydroxy-1-methylpropyl, 2-aminoethyl, 3-aminopropyl,2-methylsulfonylethyl, aminosulfonylmethyl, aminosulfonylethyl,aminosulfonylpropyl, methylaminosulfonylmethyl,methylaminosulfonylethyl, methylaminosulfonylpropyl, and the like.

[0055] “Heterocyclyl” means a saturated or unsaturated non-aromaticcyclic radical of 3 to 8 ring atoms in which one or two ring atoms areheteroatoms selected from NR^(x) {wherein each R^(x) is independentlyhydrogen, alkyl, acyl, alkylsulfonyl, aminosulfonyl,(alkylamino)sulfonyl, (dialkylamino)sulfonyl, carbamoyl,(alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl)alkyl,(alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl}, O, or S(O)_(n)(where n is an integer from 0 to 2), the remaining ring atoms being C.The heterocyclyl ring may be optionally substituted independently withone, two, or three substituents selected from alkyl, haloalkyl,heteroalkyl, halo, nitro, cyanoalkyl, hydroxy, alkoxy, amino,monoalkylamino, dialkylamino, aralkyl, —(X)_(n)—C(O)R (where X is O orNR′, n is 0 or 1, R is hydrogen, alkyl, haloalkyl, hydroxy, alkoxy,amino, monoalkylamino, dialkylamino or optionally substituted phenyl,and R′ is hydrogen or alkyl), -alkylene-C(O)R (where R is hydrogen,alkyl, haloalkyl, hydroxy, alkoxy, amino, monoalkylamino, dialkylaminoor optionally substituted phenyl) or —S(O)_(n)R^(d) (where n is aninteger from 0 to 2, and R^(d) is hydrogen (provided that n is 0),alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, amino, monoalkylamino,dialkylamino, or hydroxyalkyl). More specifically the term heterocyclylincludes, but is not limited to, tetrahydropyranyl, piperidino,N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl,3-pyrrolidino, morpholino, thiomorpholino, thiomorpholino-1-oxide,thiomorpholino-1,1-dioxide, tetrahydrothiophenyl-S,S-dioxide,pyrrolinyl, imidazolinyl, and the derivatives thereof.

[0056] “Hydroxyalkyl” means an alkyl radical as defined herein,substituted with one or more, preferably one, two or three hydroxygroups, provided that the same carbon atom does not carry more than onehydroxy group. Representative examples include, but are not limited to,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,4-hydroxybutyl, 2,3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl,2,3-dihydroxybutyl, 3,4-dihydroxybutyl and2-(hydroxymethyl)-3-hydroxypropyl, preferably 2-hydroxyethyl,2,3-dihydroxypropyl and 1-(hydroxymethyl)-2-hydroxyethyl. Accordingly,as used herein, the term “hydroxyalkyl” is used to define a subset ofheteroalkyl groups.

[0057] “Leaving group” has the meaning conventionally associated with itin synthetic organic chemistry, i.e., an atom or a group capable ofbeing displaced by a nucleophile and includes halo (such as chloro,bromo, and iodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy(e.g., acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N,O-dimethylhydroxylamino, and the like.

[0058] “Optionally substituted phenyl” means a phenyl group which isoptionally substituted with one or more substituents, preferably one,two or three, substituents preferably selected from the group consistingof alkyl, haloalkyl, hydroxyalkyl, heteroalkyl, acyl, acylamino, amino,alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl,—SO₂NR′R″ (where R′ and R″ are independently hydrogen or alkyl), alkoxy,haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halo, nitro, cyano,mercapto, methylenedioxy or ethylenedioxy. More specifically the termincludes, but is not limited to, phenyl, chlorophenyl, fluorophenyl,bromophenyl, methylphenyl, ethylphenyl, methoxyphenyl, cyanophenyl,4-nitrophenyl, 4-trifluoromethylphenyl, 4-chlorophenyl,3,4-difluorophenyl, 2,3-dichlorophenyl, 3-methyl-4-nitrophenyl,3-chloro-4-methylphenyl, 3-chloro-4-fluorophenyl or 3,4-dichlorophenyland the derivatives thereof.

[0059] “Optional” or “optionally” means that the subsequently describedevent or circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “aryl group optionally mono- ordi-substituted with an alkyl group” means that the alkyl may but neednot be present, and the description includes situations where the arylgroup is mono- or disubstituted with an alkyl group and situations wherethe aryl group is not substituted with the alkyl group.

[0060] “Pharmaceutically acceptable excipient” means an excipient thatis useful in preparing a pharmaceutical composition that is generallysafe, non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

[0061] “Pharmaceutically acceptable salt” of a compound means a saltthat is pharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

[0062] “Phenylalkyl” refers to an alkyl radical as defined herein inwhich one of the hydrogen atoms of the alkyl radical has been replacedby an optionally substituted phenyl.

[0063] “Protecting group” refers to a grouping of atoms that whenattached to a reactive group in a molecule masks, reduces or preventsthat reactivity. Examples of protecting groups can be found in T. W.Green and P. G. Futs, Protective Groups in Organic Chemistry, (Wiley,2^(nd) ed. 1991) and Harrison and Harrison et al., Compendium ofSynthetic Organic Methods, Vols. 1-8 (John Wiley and Sons, 1971-1996).Representative amino protecting groups include, formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl(Boc), trimethyl silyl (TMS), 2-trimethylsilyl-ethanesulfonyl (SES),trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (FMOC), nitro-veratryloxycarbonyl (NVOC),and the like. Representative hydroxy protecting groups include thosewhere the hydroxy group is either acylated or alkylated such as benzyl,and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers,trialkylsilyl ethers and allyl ethers.

[0064] “Treating” or “treatment” of a disease includes: (1) preventingthe disease, i.e., causing the clinical symptoms of the disease not todevelop in a mammal that may be exposed to or predisposed to the diseasebut does not yet experience or display symptoms of the disease; (2)inhibiting the disease, i.e., arresting or reducing the development ofthe disease or its clinical symptoms; or (3) relieving the disease,i.e., causing regression of the disease or its clinical symptoms.

[0065] “A therapeutically effective amount” means the amount of acompound that, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

[0066] Compounds that have the same molecular Formula but differ in thenature or sequence of bonding of their atoms or the arrangement of theiratoms in space are termed “isomers.” Isomers that differ in thearrangement of their atoms in space are termed “stereoisomers”.Stereoisomers that are not mirror images of one another are termed“diastereomers” and those that are non-superimposable mirror images ofeach other are termed “enantiomers”. When a compound has an asymmetriccenter, for example, if a carbon atom is bonded to four differentgroups, a pair of enantiomers is possible. An enantiomer can becharacterized by the absolute configuration of its asymmetric center andis described by the R- and S-sequencing rules of Cahn and Prelog, or bythe manner in which the molecule rotates the plane of polarized lightand designated as dextrorotatory or levorotatory (i.e., as (+) or(−)-isomers respectively). A chiral compound can exist as eitherindividual enantiomer or as a mixture thereof. A mixture containingequal proportions of the enantiomers is called a “racemic mixture”.

[0067] The compounds of this invention may possess one or moreasymmetric centers; such compounds can therefore be produced asindividual (R)- or (S)-stereoisomers or as mixtures thereof. Unlessindicated otherwise, the description or naming of a particular compoundin the specification and claims is intended to include both individualenantiomers and mixtures, racemic or otherwise, thereof. The methods forthe determination of stereochemistry and the separation of stereoisomersare well-known in the art (see discussion in Chapter 4 of “AdvancedOrganic Chemistry”, 4th edition J. March, John Wiley and Sons, New York,1992). In general, the nomenclature used in this Application is based onAUTONOM™, a Beilstein Institute computerized system for the generationof IUPAC systematic nomenclature. For example, a compound of Formula (I)wherein R₁ is methylene; R₂ is 4-chlorophenyl; L is C(═O); A iscyclopentyl; R₃ is hydrogen; R₄ is cyclohexyl; and D is —CH— (Example 1)is named:

[0068] Cyclohexanecarboxylic acid{2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide.

[0069] Representative compounds of Formula (I) are shown in thefollowing table. Structure M.P. (° C.) Example

1

2

2

2

2

3

PREFERRED EMBODIMENTS

[0070] While the broadest definition of this invention is set forth inthe Summary of the Invention, certain compounds of Formula (I) arepreferred.

[0071] A preferred compound of the invention is a compound of Formula(I) wherein R¹ is methylene.

[0072] Another preferred compound of the invention are compounds ofFormula (I) wherein ring A is cyclopentyl. Compounds where ring A iscyclopentyl are bind unexpectedly potently to the CCR-3 receptor. Otherpreferred compounds of the invention are compounds of Formula (I)wherein ring A is heterocyclyl (particularly tetrahydropyranyl,S,S-dioxo-tetrahydothiophenyl, tetrahydrothiophenyl or pyrrolidinyl) orcompounds of Formula (I) wherein ring A is phenyl.

[0073] A preferred compound of the invention is a compound of Formula(I) wherein R² is phenyl ring substituted with one, or two substituentsselected from alkyl, alkoxy, haloalkyl, halo, cyano or nitro; preferablymethyl, ethyl, methoxy, trifluoromethyl, chloro, fluoro or bromo; mostpreferably 4-nitrophenyl, 4-trifluoromethylphenyl, 4-chlorophenyl,3,4-difluorophenyl, 2,3-dichlorophenyl, 3-methyl-4-nitrophenyl,3-chloro-4-methylphenyl, 3-chloro-4-fluorophenyl or 3,4-dichlorophenyl.Particularly preferred are 4-chlorophenyl or 3,4-dichlorophenyl.

[0074] A preferred compound of the invention is a compound of Formula(I) wherein R³ is hydrogen or methyl, preferably hydrogen.

[0075] A preferred compound of the invention is a compound of Formula(I) wherein L is —C(═O)—, —SO₂—, —C(═O)N(R_(a))—, —C(═S)N(R_(a))— or—C(═O)O—. More preferred are compounds where L is —C(═O)—,—C(═O)N(R_(a))—, most preferably —C(═O)N(R_(a))—. In the preceding R_(a)is preferably hydrogen or methyl, most preferably hydrogen.

[0076] A preferred compound of the invention is a compound of Formula(I) wherein D is N. When D is C—R^(b), compounds that are preferred arewhere R^(b) is hydrogen.

[0077] A preferred compound of the invention is a compound of Formula(I) wherein R⁴ is alkyl, cycloalkyl, cycloalkyl alkenyl or acylalkyl;more preferably cyclohexyl, allyl, isopropyl, n-butyl, or2-(ethoxycarbonyl)ethyl.

[0078] A specific compound of Formula (I) is a compound of Formula (II):

[0079] wherein R¹-R⁴, A, D, and L have any of the values describedherein.

[0080] A specific compound of Formula (I) is a compound of Formula(III):

[0081] wherein R¹-R⁴, A, D and L have any of the values describedherein.

[0082] A specific compound of Formula (I) is a compound of formula (IV):

[0083] wherein R^(3,) R⁴, A, D and L have any of the values describedherein.

[0084] A specific compound of Formula (I) is a compound of formula (V):

[0085] wherein R⁴ and D have any of the values defined herein.

[0086] A specific compound of Formula (I) is a compound of formula (VI):

[0087] wherein R⁴ and D have any of the values defined herein.

[0088] A specific compound of Formula (I) is a compound of formula(VII):

[0089] wherein R⁴ and D have any of the values defined herein.

[0090] A specific compound of Formula (I) is a compound of formula(VIII):

[0091] wherein R⁴ and D have any of the values defined herein; and R^(x)is hydrogen, alkyl, acyl, alkylsulfonyl, aminosulfonyl,(alkylamino)sulfonyl, (dialkylamino)sulfonyl, carbamoyl,(alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl)alkyl,(alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl.

[0092] A specific compound of Formula (I) is a compound of formula (IX):

[0093] wherein R⁴ and D have any of the values defined herein.

[0094] A specific compound of Formula (I) is a compound of formula (X):

[0095] wherein R⁴ and D have any of the values defined herein.

[0096] Particularly preferred compounds of the invention are:

[0097] Cyclohexanecarboxylic acid{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide;

[0098]{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea;

[0099]1-Allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

[0100]1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;

[0101]1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

[0102]3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)propionicacid ethyl ester;

[0103] or a salt thereof.

[0104] General Utility

[0105] The compounds of the invention are CCR-3 receptor antagonists andinhibit eosinophil recruitment by CCR-3 chemokines such as RANTES,eotaxin, MCP-2, MCP-3 and MCP-4. Compounds of this invention andcompositions containing them are useful in the treatment ofeosiniphil-induced diseases such as inflammatory or allergic diseasesand including respiratory allergic diseases such as asthma, allergicrhinitis, hypersensitivity lung diseases, hypersensitivity pneumonitis,eosinophilic pneumonias (e.g., chronic eosinophilic pneumonia);inflammatory bowel diseases (e.g., Crohn's disease and ulcerativecolitis); and psoriasis and inflammatory dermatoses such as dermatitisand eczema.

[0106] Testing

[0107] The CCR-3 antagonistic activity of the compounds of thisinvention can be measured by in vitro assays such as ligand binding andchemotaxis assays as described in more detail in Examples 4, 5, and 6.In vivo activity can be assayed in the Ovalbumin induced Asthma inBalb/c Mice Model as described in more detail in Example 7.

[0108] Administration and Pharmaceutical Composition

[0109] In general, the compounds of this invention can be administeredin a therapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. The actualamount of the compound of this invention, i.e., the active ingredient,will depend upon numerous factors such as the severity of the disease tobe treated, the age and relative health of the subject, the potency ofthe compound used, the route and form of administration, and otherfactors.

[0110] Therapeutically effective amounts of compounds of Formula (I) mayrange from approximately 0.01-20 mg per kilogram body weight of therecipient per day; preferably about 0.1-10 mg/kg/day. Thus, foradministration to a 70 kg person, the dosage range would most preferablybe about 7 mg to 0.7 g per day.

[0111] In general, compounds of this invention will be administered aspharmaceutical compositions by any one of the following routes: oral,transdermal, inhalation (e.g., intranasal or oral inhalation) orparenteral (e.g., intramuscular, intravenous or subcutaneous)administration. A preferred manner of administration is oral using aconvenient daily dosage regimen which can be adjusted according to thedegree of affliction. Compositions can take the form of tablets, pills,capsules, semisolids, powders, sustained release formulations,solutions, suspensions, liposomes, elixirs, or any other appropriatecompositions. Another preferred manner for administering compounds ofthis invention is inhalation. This is an effective means for deliveringa therapeutic agent directly to the respiratory tract for the treatmentof diseases such as asthma and other similar or related respiratorytract disorders (see U.S. Pat. No. 5,607,915).

[0112] The choice of formulation depends on various factors such as themode of drug administration and the bioavailability of the drugsubstance. For delivery via inhalation the compound can be formulated asliquid solutions or suspensions, aerosol propellants or dry powder andloaded into a suitable dispenser for administration. There are threetypes of pharmaceutical inhalation devices—nebulizer inhalers,metered-dose inhalers (MDI) and dry powder inhalers (DPI). Nebulizerdevices produce a stream of high velocity air that causes thetherapeutic agents (which has been formulated in a liquid form) to sprayas a mist which is carried into the patient's respiratory tract. MDI'stypically have the Formulation packaged with a compressed gas. Uponactuation, the device discharges a measured amount of therapeutic agentby compressed gas, thus affording a reliable method of administering aset amount of agent. DPI's administer therapeutic agents in the form ofa free flowing powder that can be dispersed in the patient's inspiratoryair-stream during breathing by the device. In order to achieve a freeflowing powder, the therapeutic agent is formulated with an excipient,such as lactose. A measured amount of the therapeutic is stored in acapsule form and is dispensed to the patient with each actuation.Recently, pharmaceutical formulations have been developed especially fordrugs that show poor bioavailability based upon the principle thatbioavailability can be increased by increasing the surface area i.e.,decreasing particle size. For example, U.S. Pat. No. 4,107,288 describesa pharmaceutical formulation having particles in the size range from 10to 1,000 nm in which the active material is supported on a crosslinkedmatrix of macromolecules. U.S. Pat. No. 5,145,684 describes theproduction of a pharmaceutical formulation in which the drug substanceis pulverized to nanoparticles (average particle size of 400 nm) in thepresence of a surface modifier and then dispersed in a liquid medium togive a pharmaceutical formulation that exhibits remarkably highbioavailability.

[0113] The compositions are comprised of in general, a compound ofFormula (I) in combination with at least one pharmaceutically acceptableexcipient. Acceptable excipients are non-toxic, aid administration, anddo not adversely affect the therapeutic benefit of the compound ofFormula (I). Such excipient may be any solid, liquid, semi-solid or, inthe case of an aerosol composition, gaseous excipient that is generallyavailable to one of skill in the art.

[0114] Solid pharmaceutical excipients include starch, cellulose, talc,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, magnesium stearate, sodium stearate, glycerol monostearate, sodiumchloride, dried skim milk and the like. Liquid and semisolid excipientsmay be selected from glycerol, propylene glycol, water, ethanol andvarious oils, including those of petroleum, animal, vegetable orsynthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesameoil, etc. Preferred liquid carriers, particularly for injectablesolutions, include water, saline, aqueous dextrose, and glycols.

[0115] Compressed gases may be used to disperse a compound of thisinvention in aerosol form. Inert gases suitable for this purpose arenitrogen, carbon dioxide, etc.

[0116] For liposomal formulations of the drug for parenteral or oraldelivery the drug and the lipids are dissolved in a suitable organicsolvent e.g. tert-butanol, cyclohexane (1% ethanol). The solution islyophilized and the lipid mixture is suspended in an aqueous buffer andallowed to form a liposome. If necessary, the liposome size can bereduced by sonication. (see, Frank Szoka, Jr. and DemetriosPapahadjopoulos, “Comparative Properties and Methods of Preparation ofLipid Vesicles (Liposomes)”, Ann. Rev. Biophys. Bioeng., 9:467-508(1980), and D. D. Lasic, “Novel Applications of Liposomes”, Trends inBiotech., 16:467-608, (1998)).

[0117] Other suitable pharmaceutical excipients and their formulationsare described in Remington's Pharmaceutical Sciences, edited by E. W.Martin (Mack Publishing Company, 18th ed., 1990).

[0118] The level of the compound in a formulation can vary within thefull range employed by those skilled in the art. Typically, theformulation will contain, on a weight percent (wt %) basis, from about0.01-99.99 wt % of a compound of Formula (I) based on the totalformulation, with the balance being one or more suitable pharmaceuticalexcipients. Preferably, the compound is present at a level of about 1-80wt %. Representative pharmaceutical formulations containing a compoundof Formula (I) are described in Example 4.

[0119] Synthesis of Compounds of Formula (I)

[0120] The compounds of the present invention can be prepared in anumber of ways known to one skilled in the art. Preferred methodsinclude, but are not limited to, the general synthetic proceduresdescribed below.

[0121] The starting materials and reagents used in preparing thesecompounds are either available from commercial suppliers such as AldrichChemical Co., (Milwaukee, Wis., USA), Bachem (Torrance, Calif., USA),Enika-Chemie, or Sigma (St. Louis, Mo., USA), Maybridge (Dist: RyanScientific, P.O. Box 6496, Columbia, S.C. 92960), Bionet Research Ltd.,(Cornwall PL32 9QZ, UK), Menai Organics Ltd., (Gwynedd, N. Wales, UK),Butt Park Ltd., (Dist. Interchim, Montlucon Cedex, France) or areprepared by methods known to those skilled in the art followingprocedures set forth in references such as Fieser and Fieser's Reagentsfor Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd'sChemistry of Carbon Compounds, Volumes 1-5 and Supplementals (ElsevierScience Publishers, 1989), Organic Reactions, Volumes 1-40 (John Wileyand Sons, 1991), March's Advanced Organic Chemistry, (John Wiley andSons, 1992), and Larock's Comprehensive Organic Transformations (VCHPublishers Inc., 1989). These schemes are merely illustrative of somemethods by which the compounds of this invention can be synthesized, andvarious modifications to these schemes can be made and will be suggestedto one skilled in the art having referred to this disclosure.

[0122] The starting materials and the intermediates of the reaction maybe isolated and purified if desired using conventional techniques,including but not limited to filtration, distillation, crystallization,chromatography, and the like. Such materials may be characterized usingconventional means, including physical constants and spectral data.

[0123] Compounds of Formula (I) are generally prepared from theprecursor amine of Formula (Ia) as shown below.

[0124] Preparation of compounds of Formula (Ia) and their conversion tocompounds of Formula I is illustrated in the following Schemes 1-8.

[0125] Schemes 1-5 show methods of preparing compounds of Formula Iahaving different rings A. Specific exemplification is provided for R¹-R²being 4-chlorobenzyl in Preparations 1-6. Preparation of analogouscompounds where R¹ and R² vary within the full scope of the Summary ofthe Invention may be readily prepared by one of skill in the art inlight of this specification and incorporated references.

General Procedure A: (Amine Alkylation with Epoxides)

[0126] A 0.5-1.5 M solution of the amine, R₂NH (1 equiv), and thespecified epoxide, 3a (1.1-10 equiv) in EtOH is stirred at 80-95° C. for2-4.5 d, allowed to cool to room temperature, and concentrated. Thecrude amino alcohol is purified by chromatography or recrystallization.

General Procedure B: (Amine Formation Using Methanesulfonyl Chloride andAmmonium Hydroxide)

[0127] A 0.2-0.3 M solution of the amino alcohol (1 equiv) in CH₂Cl₂ at0° C. is treated successively with Et₃N (2 equiv) and MeSO₂Cl (2 equiv),stirred at 0° C. for 1-2 hours, and partitioned between CH₂Cl₂ and10-15% NH₄OH. The aqueous phase is extracted with CH₂Cl₂ and theextracts are dried and concentrated. A 0.13M solution of the residue in2.5:1 dioxane:28-30 wt % NH₄OH is stirred at 70-80° C. 2.5-18 hours,allowed to cool to room temperature, and concentrated. The residue ispartitioned between EtOAc and 1 N NaOH, the aqueous phase is extractedwith EtOAc, and the extracts are washed with brine, dried andconcentrated. The crude product is purified by chromatography or usedwithout further purification.

[0128] Schemes 6 and 7 show preparation of compounds of Formula Ia wherering A is substituted. Scheme 6 shows preparation of compounds ofFormula Ia with a substituted cyclopentyl ring A. Scheme 7 showspreparation of compounds of Formula Ia with a substituted pyrrolidinering A by treatment of the unsubstituted pyrrolidine 7a (R═H) with theappropriate reagent to produce the substituted pyrrolidine 7b.

[0129] Schemes 8 and 9 show methods of converting compounds of Formula(Ia) to compounds of Formula (I) where L and A are varied.

[0130] Scheme 8

General Procedure C: (Urea Formation Using Isocyanates)

[0131] A 0.1-0.6 M solution of the amine (1 equiv) in CH₂Cl₂ or CH₂Cl₂and DMF at 0-20° C. is treated with the specified isocyanate (1.1-2equiv), stirred for 0.5-1.5 hours, and partitioned between CH₂Cl₂ andsaturated NaHCO₃. The aqueous phase is extracted with CH₂Cl₂ and theextracts are dried and concentrated. The crude urea is purified bycolumn chromatography or preparative TLC or used in the next stepwithout further purification.

General Procedure D: (Urea Formation Using Isocyanates)

[0132] A 0.1-0.6 M solution of the amine (1 equiv) in CH₂Cl₂ or CH₂Cl₂and DMF at 0-20° C. is treated with the specified isocyanate (1.1-2equiv), stirred for 0.5-1.5 hours, and partitioned between CH₂Cl₂ andsaturated NaHCO₃. The aqueous phase is extracted with CH₂Cl₂ and theextracts are dried and concentrated. The crude urea is purified bycolumn chromatography or preparative TLC or used in the next stepwithout further purification. A solution of the free base in CH₂Cl₂ istreated with 1 N HCl in Et₂O and concentrated to give the hydrochloridesalt?

General Procedure E: (Amide Formation Using 1-Hydroxybenzotriazole and1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride)

[0133] A 0.1-0.4 M solution of the amine (1 equiv) and the specifiedcarboxylic acid (1.2-1.5 equiv) in CH₂Cl₂ at 0° C. is treatedsuccessively with 1-hydroxybenzotriazole hydrate (HOBt) (0.2-0.5 equiv)and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (DEC)(1.3-2 equiv), stirred at 0-20° C. for 2-72 hours, and partitionedbetween CH₂Cl₂ and saturated NaHCO₃. The aqueous phase is extracted withCH₂Cl₂ and the extracts are dried and concentrated. The crude amide ispurified by column chromatography and/or preparative TLC.

General Procedure F: (Amide Formation Using 1-Hydroxybenzotriazole and1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride)

[0134] A 0.1-0.4 M solution of the amine (1 equiv) and the specifiedcarboxylic acid (1.2-1.5 equiv) in CH₂Cl₂ at 0° C. is treatedsuccessively with 1-hydroxybenzotriazole hydrate (HOBt) (0.2-0.5 equiv)and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (DEC)(1.3-2 equiv), stirred at 0-20 C. for 2-72 hours, and partitionedbetween CH₂Cl₂ and saturated NaHCO₃. The aqueous phase is extracted withCH₂Cl₂ and the extracts are dried and concentrated. The crude amide ispurified by column chromatography and/or preparative TLC. A solution ofthe free base in CH₂Cl₂ is treated with 1 N HCl in Et₂O and concentratedto provide the hydrochloride salt.

[0135] Scheme 9 and following procedures G-O describe the variousmethods used to convert compounds of Formula Ia to compounds of FormulaI where L is varied.

General Procedure G (Parallel Synthesis of Sulfonamides)

[0136] A mixture of the requisite amine Ia (1 equiv), the appropriatesulfonyl chloride (1.5 equiv), and Amberlite IRA67 (2 equiv) in CH₂Cl₂(2 mL) was rotated overnight. The mixture was treated with PS-trisamine(1.2 equiv) (Argonaut Technologies Inc., San Carlos, Calif., USA) androtated overnight. The solid was collected by filtration and washed withCH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate was concentrated to give theproduct.

General Procedure H (Parallel Synthesis of Amides from Acid Chlorides)

[0137] A mixture of the requisite amine Ia (1 equiv), the appropriateacid chloride (1.5 equiv), and Amberlite IRA67 (2 equiv) in CH₂Cl₂ (2mL) was rotated overnight. The mixture was treated with PS-trisamine(1.2 equiv) and MP-carbonate (2 equiv) (Argonaut Technologies, SanCarlos, Calif.) and rotated overnight. The solid was collected byfiltration and washed with CH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate wasconcentrated to give the product.

General Procedure I (Parallel Synthesis of Amides from Carboxylic Acids)

[0138] A mixture of the requisite amine Ia (1 equiv), the appropriatecarboxylic acid (1.5 equiv), and PS-carobodiimide (2 equiv) (ArgonautTechnologies Inc., San Carlos, Calif., USA) in CH₂Cl₂ (2 mL) was rotatedovernight. The mixture was treated with MP-carbonate (2 equiv) androtated overnight. The solid was collected by filtration and washed withCH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate was concentrated to give theproduct.

General Procedure J (Parallel Synthesis of Ureas from Isocyanates andPurification by Parallel Chromatography)

[0139] A mixture of the requisite amine Ia (1 equiv) and the appropriateisocyanate (1.2 equiv) in CH₂Cl₂ (2 mL) was stirred overnight. Themixture was concentrated to give the crude product, which was purifiedby parallel chromatography using a step gradient (2.5% MeOH/CH₂Cl₂, 10%MeOH/CH₂Cl₂).

General Procedure K (Parallel Synthesis of Ureas from Isocyanates andPurification by Catch and Release Scavenger)

[0140] A mixture of the requisite amine Ia (1 equiv) and the appropriateisocyanate (1.2 equiv) in CH₂Cl₂ (2 mL) was stirred overnight. Themixture was treated with MP-TsOH and rotated for 3 h. The solid wascollected by filtration and washed with CH₂Cl₂, MeOH, and CH₂Cl₂. Thesolid was rotated with 2 M NH₃ in MeOH for 2 h. The solid was collectedby filtration and washed with CH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate wasconcentrated to give the purified product.

General Procedure L (Parallel Synthesis of Ureas from Anilines usingPhoxime Resin)

[0141] A mixture of the appropriate aniline (3 equiv) and Phoxime resin(1 equiv) in CH₂Cl₂ (2 mL) was rotated for 3 h. If the aniline had notdissolved, triethylamine (3.5 equiv) was added. The mixture was rotatedovernight. The solid was collected by filtration and washed with CH₂Cl₂,MeOH, CH₂Cl₂, MeOH, and CH₂Cl₂. A mixture of the solid and the requisiteamine Ia (1.1 equiv) in CH₂Cl₂ (0.5 mL) and toluene (1.5 mL) were heatedat 80° C. with shaking overnight and allowed to cool to roomtemperature. The solid was collected by filtration and washed withCH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate was concentrated to give theproduct.

General Procedure M (Parallel Synthesis of Ureas from Anilines usingTriphosgene)

[0142] A mixture of the appropriate aniline (1.2 equiv), triphosgene(0.4 equiv), and triethylamine (1.4 equiv) in CH₂Cl₂ was heated at 35°C. for 1 h. After cooling to room temperature, the requisite amine Ia (1equiv) was added. The mixture was stirred overnight, washed with H₂O andbrine, passed through Na₂SO₄, and concentrated to give crude productwhich was purified by parallel chromatography.

General Procedure N (Parallel Synthesis of Thioureas fromThioisocyanates)

[0143] A mixture of the requisite amine Ia (1 equiv) and the appropriatethioisocyanate (1.2 equiv) in CH₂Cl₂ (2 mL) was stirred overnight. Themixture was treated with MP-TsOH and rotated for 3 h. The solid wascollected by filtration and washed with CH₂Cl₂, MeOH, and CH₂Cl₂. Thesolid was rotated with 2 M NH₃ in MeOH for 2 h. The solid was collectedby filtration and washed with CH₂Cl₂, MeOH, and CH₂Cl₂. The filtrate wasconcentrated to give the purified product.

General Procedure O (Parallel Synthesis of Carbamates)

[0144] A mixture of the requisite amine Ia (1 equiv) and the appropriatesuccinimide (1.5 equiv) in CH₂Cl₂ (2 mL) was stirred overnight. If thereaction was not complete, it was heated at 38° C. for 1 hour. Themixture was washed with H₂O and brine, passed through Na₂SO₄ andconcentrated to give the crude product which was purified via parallelpurification (step gradient 5% MeOH/CH₂Cl₂, 10% MeOH/CH₂Cl₂).

Experimental Section

[0145] General

[0146] Unless otherwise noted, all non-aqueous reactions were run undera nitrogen atmosphere and Na₂SO₄ was used to dry all organic layers.Purifications were typically carried out by flash chromatography onsilica gel (230-400 mesh) or preparative TLC on Uniplate Silica Gel GFPLC Plates (20×20 cm, 1000 microns) from Analtech, Inc., Newark, Del.Alumina used was basic with 6 wt % H₂O (Brockmann III). Melting pointstaken in capillary tubes are uncorrected. IR spectra were determined inKBr. NMR spectra were run in CDCl₃, unless otherwise indicated. ¹H NMRspectra were recorded on 300 MHz instruments and ¹³C NMR spectra wererecorded at 75.5 MHz. Mass spectral analyses were accomplished usingelectrospray ionization. Analytical reverse-phase HPLC was performed onShimadzu system equipped with a diode array spectrometer (range 190-300nm; Hewlett Packard). The stationary phase was a Zorbax SB-Phenyl RapidResolution column (4.6 mm×50 mm; Hewlett Packard), mobile phase A was0.1% trifluoroacetic acid, and mobile phase B was CH₃CN. A flow rate of2.5 mL/min with a linear gradient of 20-55% B in 5 min and then 55-20% Bin 5 min was employed. All parallel synthesis reactions were run insealed tubes that were vented prior to being rotated overnight.Amberlite IRA67 (Aldrich Chemical Co., Milwaukee, Wis., USA) was washedconsecutively with CH₂Cl₂, MeOH, CH₂Cl₂, MeOH, CH₂Cl₂ and then driedunder vacuum prior to use. All products derived from parallel synthesisreactions were characterized via HPLC-MS.

EXAMPLES

[0147] The following Preparations and Examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof.

[0148] The following Preparations (1-7) are useful for preparingsynthetic intermediates that can be used to prepare compounds of theinvention, as described in the Schemes and Examples.

Preparation 1: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

[0149]

Step A: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexanol

[0150]

[0151] Following Procedure A, 4-(4-chlorobenzyl)-piperidine (seePreparation 7) (52 mg, 0.25 mmol) was alkylated with7-oxa-bicyclo[4.1.0]heptane (0.25 mL, 2.5 mmol) in EtOH (0.5 mL) at 80°C. for 3 d. Chromatography of the crude product with 90:9.5:0.5-80:19:1CH₂Cl₂:MeOH:NH₄OH gave the product (68 mg, 88%) as a tan oil whichsolidified upon standing as a cream solid: mp 100-101.3° C.; IR 3379,2929 cm⁻¹; ¹H NMR δ 1.05-1.76 (m, 12H), 2.02 (dt, J=2.4,11.6 Hz, 1H),2.06-2.20 (m, 2H), 2.49 (d, J=7.0 Hz, 2H), 2.51-2.64 (m, 2H), 2.79 (m,1H), 3.34 (m, 1H), 4.05 (m, 1H), 7.06 (m, 2H), 7.24 (m, 2H); MS m/z 308(M+H)⁺. Anal. (C₁₈H₂₆ClNO) C, H, N.

Step B: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

[0152]

[0153] A solution of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexanol (390 mg,1.27 mmol) in CH₂Cl₂ (6 mL) at 0° C. was treated successively with Et₃N(350 μL, 2.53 mmol) and MeSO₂Cl (194 μL, 2.53 mmol), stirred at 0° C.for 2 hours, and partitioned between CH₂Cl₂ and 10% NH₄OH. The aqueousphase was extracted with CH₂Cl₂ and the extracts were washed with brine,dried and concentrated. A solution of the residue in THF (3 mL) and28-30 wt % NH₄OH (1.2 mL) was stirred at 70° C. for 24 hours, allowed tocool to room temperature, and partitioned between EtOAc and 1 N NaOH.The aqueous phase was extracted with EtOAc and the extracts were washedwith brine, dried and concentrated. Chromatography of the residue onalumina with 1:3 EtOAc:MeOH to 100% MeOH and a subsequent chromatographyon alumina with 20:1 hexanes:EtOAc to 100% EtOAc followed by 3:1EtOAc:MeOH to 100% MeOH gave the product (260 mg, 67%) as a tan oilwhich solidified upon standing: mp 69.1-70.4° C.; ¹H NMR δ 1.03-1.34 (m,6H), 1.37-1.52 (m, 1H), 1.57-1.77 (m, 5H), 1.92-2.05 (m, 3H), 2.48 (d,J=7.0 Hz, 2H), 2.45-2.64 (m, 3H), 2.73 (m, 1H), 7.06 (m, 2H), 7.23 (m,2H); MS m/z 307 (M+H)⁺.

Preparation 2: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

[0154]

Step A: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentanol

[0155]

[0156] Following General Procedure A, a solution of4-(4-chlorobenzyl)-piperidine (17.86 g, 85.05 mmol) and6-oxa-bicyclo[3.1.0]hexane (50 g, 0.6 mol) in EtOH (170 mL) was stirredat 95° C. for 40 hours, allowed to cool to room temperature, andconcentrated. The residue was crystallized in hot CH₂Cl₂ (80 mL), thecrystallization mixture was concentrated to half the volume, and kept at0° C. overnight and filtered, and the precipitate was rinsed with coldhexanes to give the product (18.2 g, 73%) as a tan solid. The motherliquors were concentrated to half the volume, diluted with CH₂Cl₂ andkept at −10° C. for 1 hour, and the precipitate was rinsed with coldCH₂Cl₂ and hexanes to give additional product (1.8 g, 7%) as a tansolid: mp 104.1-105.5° C.; IR 3436, 2928 cm⁻¹; ¹H NMR δ 1.19-1.75 (m,8H), 1.81-1.99 (m, 4H), 2.06 (dt, J=2.5,11.7 Hz, 1H), 2.47 (m, 1H), 2.50(d, J=7.0 Hz, 2H), 2.90 (m, 1H), 3.07 (m, 1H), 4.10 (m, 1H), 7.06 (m,2H), 7.23 (m, 2H); ¹³C NMR δ 21.63, 27.35, 32.01, 32.15, 34.31, 37.87,42.47, 50.47, 52.97, 75.15, 75.22, 128.27, 130.43, 131.55, 139.04; MSm/z 294 (M+H)⁺. Anal. (C₁₇H₂₄ClNO.0.1H₂O) C, H, N.

Step B: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

[0157]

[0158] Following General Procedure B, a solution of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentanol (205 mg,0.697 mmol) in CH₂Cl₂ (2.8 mL) at 0° C. was treated successively withEt₃N (190 μL, 1.4 mmol) and MeSO₂Cl (110 μL, 1.4 mmol), stirred at 0° C.for 1 hour, and partitioned between CH₂Cl₂ and 10% NH₄OH. The aqueousphase was extracted with CH₂Cl₂ and the extracts were dried andconcentrated to give 220 mg of an oil. A solution of the residue (110mg) in dioxane (2 mL) and 28-30 wt % NH₄OH (0.8 mL) was stirred at70-80° C. overnight, allowed to cool to room temperature, andconcentrated. The residue was partitioned between EtOAc and 1 N NaOH,the aqueous phase was extracted with EtOAc, and the extracts were washedwith brine, dried and concentrated. Chromatography of the residue onalumina with 10:1 hexanes:EtOAc to 100% EtOAc followed by95:4.75:0.25-60:38:2 CH₂Cl₂:MeOH:NH₄OH gave the product (87 mg, 85%) asan oil: ¹H NMR δ 1.18-1.71 (m, 9H), 1.76-2.00 (m, 3H), 2.07 (dt,J=2.4,11.5 Hz, 1H), 2.31 (m, 1H), 2.50 (d, J=6.9 Hz, 2H), 2.86-2.99 (m,2H), 3.19 (m, 1H), 7.06 (m, 2H), 7.23 (m, 2H); MS m/z 293.2 (M+H)⁺.

Preparation 3: Preparation of(±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

[0159]

Step A: Preparation of (±)-trans-4-nitro-benzenesulfonic Acid2-azido-cyclopentyl Ester

[0160]

[0161] A solution of (±)-trans-2-azido-cyclopentanol (1.27 g, 10.0 mmol)(Zhang, Z. da; Scheffold, R. Helv. Chim. Acta 1993, 76, 2602) in CH₂Cl₂(14 mL) at 0° C. was treated successively with pyridine (0.88 mL, 10.9mmol) and 4-nitro-benzenesulfonyl chloride (2.22 g, 10.0 mmol) andallowed to warm to room temperature slowly. The reaction was stirred for4 d, during which additional pyridine (0.9 mL, 11 mmol) and4-nitro-benzenesulfonic acid (2.2 g, 10 mmol) was added, and partitionedbetween CH₂Cl₂ and 1 N HCl. The aqueous phase was extracted with CH₂Cl₂and the extracts were washed with saturated NaHCO₃, dried andconcentrated. Chromatography of the residue with 10:1-4:1 hexanes:EtOAcgave the product (2.63 g, 84%) as a yellow oil: ¹H NMR δ 1.61-1.90 (m,4H), 2.00-2.16 (m, 2H), 3.96 (m, 1H), 4.72 (m, 1H), 8.14 (m, 2H), 8.43(m, 2H).

Step B: Preparation of(±)-cis-1-(2-azido-cyclopentyl)-4-(4-chlorobenzyl)-piperidine

[0162]

[0163] A murky solution of (±)-trans-4-nitro-benzenesulfonic acid2-azido-cyclopentyl ester (630 mg, 2.0 mmol),4-(4-chlorobenzyl)-piperidine (420 mg, 2.0 mmol), and Et₃N (280 μL, 2.0mmol) in CH₃CN (4 mL) was stirred at room temperature for 10 d and 65°C. for 2 d, allowed to cool to room temperature, and concentrated. Theresidue was partitioned between CH₂Cl₂ and 1 N NaOH, the aqueous phasewas extracted with CH₂Cl₂ and the extracts were dried and concentrated.Chromatography of the residue with 20:1-1:1 hexanes:EtOAc followed bychromatography with 100% CH₂Cl₂to 95:4.75:0.25 CH₂Cl₂:MeOH:NH₄OH gavethe product (145 mg, 22%) as a tan oil: ¹H NMR δ 1.32-1.90 (m, 13H),2.33 (m, 1H), 2.49 (d, J=6.4 Hz, 2H), 2.96 (m, 1H), 3.06 (m, 1H), 4.04(t, J=4.0 Hz, 1H), 7.06 (m, 2H), 7.23 (m, 2H); MS m/z 319.2 (M−H)⁻.

Step C: Preparation of(±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

[0164]

[0165] A solution of(±)-cis-1-(2-azido-cyclopentyl)-4-(4-chlorobenzyl)-piperidine (210 mg,0.65 mmol) in THF (2.5 mL) was treated successively with PPh₃ (514 mg,1.96 mmol) and H₂O (141 μL, 7.83 mmol), refluxed for 3.5 hours, allowedto cool to room temperature, and concentrated. Chromatography of theresidue with 90:9.5:0.5-75:23.75:1.25 CH₂Cl₂:MeOH:NH₄OH gave the product(183 mg, 95%) as a colorless oil which solidified upon standing to acream solid: mp 69.6-71.3° C.; ¹H NMR δ 1.20-1.35 (m, 2H), 1.43-1.93 (m,11H), 2.17 (m, 1H), 2.49 (d, J=6.9 Hz, 2H), 2.89-3.02 (m, 2H), 3.34 (t,J=4.4 Hz, 1H), 7.06 (m, 2H), 7.23 (m, 2H); ¹³C NMR δ 20.72, 27.08,32.48, 32.61, 38.32, 42.95, 52.14, 53.09, 53.61, 71.49, 128.63, 130.80,131.88, 139.58; MS m/z 293.2 (M+H)⁺.

Preparation 4: Preparation of(±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

[0166]

Step A: Preparation of (±)-trans-4-nitro-benzenesulfonic Acid2-azido-cyclohexyl Ester

[0167]

[0168] A solution of (±)-trans-2-azidocyclohexan-1-ol (11.3 g, 80.0mmol) (Zhang, Z. da; Scheffold, R. Helv. Chim. Acta 1993, 76, 2602] inCH₂Cl₂ (110 mL) at 0° C. was treated successively with pyridine (14.2mL, 176 mmol) and 4-nitro-benzenesulfonyl chloride (35.6 g, 160 mmol),allowed to warm to room temperature slowly, stirred for 4 d, andpartitioned between CH₂Cl₂ and 1 N HCl. The aqueous phase was extractedwith CH₂Cl₂ and the extracts were washed with saturated NaHCO₃, driedand concentrated. Chromatography of the residue with 10:1-1:1hexanes:EtOAc gave the product (19 g, 72%) as a cream solid: ¹H NMR δ1.19-1.39 (m, 3H), 1.53-1.82 (m, 3H), 2.00-2.10 (m, 1H), 2.26 (m, 1H),3.36 (m, 1H), 4.35 (ddd, J=4.7, 9.2, 10.8 Hz, 1H), 8.17 (m, 2H), 8.41(m, 2H).

Step B: Preparation of(±)-cis-1-(2-azido-cyclohexyl)-4-(4-chlorobenzyl)-piperidine

[0169]

[0170] A murky solution of (±)-trans-4-nitro-benzenesulfonic acid2-azido-cyclohexyl ester (1.77 g, 5.41 mmol),4-(4-chlorobenzyl)-piperidine (1.14 g, 5.43 mmol), and Et₃N (0.75 mL,5.4 mmol) in CH₃CN (11.2 mL) was stirred at room temperature for 17hours, 65° C. for 31 hours, and 80° C. for 5 d, allowed to cool to roomtemperature, and concentrated. The residue was partitioned betweenCH₂Cl₂ and 1 N NaOH, the aqueous phase was extracted with CH₂Cl₂ and theextracts were dried and concentrated. Chromatography of the residue with98:1.9:0.1-95:4.75:0.25 CH₂Cl₂:MeOH:NH₄OH to 100% MeOH and subsequentchromatography with 10:1 hexanes:EtOAc to 100% EtOAc followed by 95:5EtOAc:MeOH gave, in order of elution, starting(±)-trans-4-nitro-benzenesulfonic acid 2-azidocyclohexyl ester (1.2 g,68%), desired product (155 mg, 9%), and starting4-(4-chlorobenzyl)-piperidine (810 mg, 71%). Product: ¹H NMR δ 1.19-1.81(m, 12H), 1.92-2.08 (m, 3H), 2.22 (m, 1H), 2.48 (d, J=7.0 Hz, 2H), 3.02(m, 2H), 4.05 (m, 1H), 7.06 (m, 2H), 7.23 (m, 2H); MS m/z 333.2 (M+H)⁺.

Step C: Preparation of(±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

[0171]

[0172] A solution of(±)-cis-1-(2-azido-cyclohexyl)-4-(4-chlorobenzyl)-piperidine (155 mg,0.463 mmol) in THF (1.8 mL) was treated successively with PPh₃ (364 mg,1.39 mmol) and H₂O (141 μL, 5.56 mmol), refluxed for 3 hours, allowed tocool to room temperature, and concentrated. Chromatography of theresidue with 95:4.75:0.25-75:23.75:1.25 CH₂Cl₂:MeOH:NH₄OH gave theproduct (121 mg, 85%) as a cream solid: ¹H NMR δ 1.14-1.93 (m, 15H),1.96 (dt, J=11.8, 3.5 Hz, 1H), 2.48 (d, J=7.0 Hz, 2H), 3.03-3.13 (m,2H), 3.30 (m, 1H), 7.06 (m, 2H), 7.23 (m, 2H); MS m/z 307.2 (M+H)⁺.

Preparation 5: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

[0173]

Step A: Preparation of(±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone

[0174]

[0175] 1,2-Bis(trimethylsilyloxy)cyclobutene (5.0 g, 22 mmol) at 0° C.under Ar was treated dropwise during 15 min with a solution of4-(4-chlorobenzyl)-piperidine (4.56 g, 21.7 mmol) in MeOH (10.9 mL) andallowed to warm to room temperature. The reaction was stirred over aperiod of 5 hours, during which additional1,2-bis(trimethylsilyloxy)cyclobutene (0.99 g, 4.3 mmol) was added, andconcentrated. Chromatography of the residue with 95:4.75:0.25CH₂Cl₂:MeOH:NH₄OH gave the product (4.8 g, 80%) as a yellow oil: ¹H NMRδ 1.20-1.35 (m, 2H), 1.43-1.64 (m, 3H), 1.93-2.18 (m, 4H), 2.49 (d,J=6.9 Hz, 2H), 2.64-2.91 (m, 3H), 3.14 (m, 1H), 3.90 (m, 1H), 7.05 (m,2H), 7.23 (m, 2H); MS m/z 278.1 (M+H)⁺.

Step B: Preparation of(±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime

[0176]

[0177] A solution of(±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone (1.74 g, 6.26mmol) and MeONH₂.HCl (2.63 g, 31.3 mmol) in MeOH (20 mL) was stirred at65° C. under Ar for 3 hours, allowed to cool to room temperature, andconcentrated. The residue was partitioned between CH₂Cl₂ and saturatedNaHCO₃, the aqueous phase was extracted with CH₂Cl₂, and the extractswere dried and concentrated. Chromatography of the residue with95:4.75:0.25 CH₂Cl₂:MeOH:NH₄OH gave the product (1.5 g, 78%) as a brownoil and predominantly one stereoisomer: ¹H NMR δ 1.05-1.65 (m, 4.5H),1.92-2.11 (m, 4H), 2.45-2.65 (m, 3H), 2.73-2.96 (m, 2H), 3.22 (m, 1H),3.73 (m, 1H), 3.82 (m, 3H), 4.57 (m, 0.5H), 7.06 (m, 2H), 7.23 (m, 2H);MS m/z 307.1 (M+H)⁺.

Step C: Preparation of(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

[0178]

[0179] A mixture of NaBH₄ (604 mg, 16.0 mmol) in THF (13 mL) under Arwas treated dropwise with trifluoroacetic acid (1.23 mL, 16.0 mmol),stirred for 5 min, treated dropwise with a solution of(±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime(985 mg, 3.21 mmol) in THF (35 mL), and stirred at room temperature for5 h. The mixture was treated carefully with 6 N HCl (1.5 mL) until thepH ˜2, stirred for 10 min, basified with 8 N NaOH until the pH ˜10, andpartitioned between EtOAc and 1 N NaOH. The aqueous phase was extractedwith EtOAc and the extracts were washed with brine, dried (Na₂SO₄) andconcentrated. A solution of the residue in MeOH (30 mL) and 1 N HCl (3mL) was stirred at 50° C. for 1 h and at 75° C. for 5 hours, allowed tocool to room temperature, and concentrated. The residue was partitionedbetween CH₂Cl₂ and 1 N NaOH, the aqueous phase was extracted with CH₂Cl₂and the extracts were dried and concentrated. Chromatography of theresidue on alumina with 10:1 hexanes:EtOAc to 100% EtOAc followed by98:1.9:0.1-90:9.5:0.5 CH₂Cl₂:MeOH:NH₄OH gave 400 mg of the product (80%pure by ¹H NMR) as a yellow oil which was used without furtherpurification: ¹H NMR δ 1.19-1.90 (m, 9H), 2.11 (m, 1H), 2.28 (m, 1H),2.44-2.59 (m, 3H), 2.80 (m, 1H), 3.05 (m, 1H), 3.22 (m, 1H), 7.06 (m,2H), 7.23 (m, 2H); MS m/z 279.2 (M+H)⁺.

Preparation 6: Preparation of(±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

[0180]

[0181] A solution of(±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime(438 mg, 1.43 mmol) in THF (13 mL) under Ar was treated dropwise with 1M BH₃.THF complex in THF (8.6 mL, 8.6 mmol) and stirred at roomtemperature for 3 h and at 75° C. for 20 h. The reaction was cooled to0° C. and treated carefully with 6 N HCl (1 mL) until the pH ˜2. The THFwas evaporated and a solution of the residue in EtOH (9 mL) and 6 N HCl(1 mL) was stirred at 75° C. for 1 h. It was then allowed to cool toroom temperature, basified with 8 N NaOH (4 mL) until the pH ˜10,diluted with H₂O (5 mL) to dissolve the resulting white precipitate, andconcentrated. The residue was partitioned between CH₂Cl₂ and 1 N NaOH,the aqueous phase was extracted with CH₂Cl₂, and the extracts were driedand concentrated. Chromatography of the residue with 90:9.5:0.5-60:38:2CH₂Cl₂:MeOH:NH₄OH gave, in order of elution, 70 mg of the desiredproduct (80% pure by ¹H NMR) as a colorless oil which was used withoutfurther purification, 48 mg (12%) of pure desired product as a colorlessoil, and 125 mg of a mixture of desired product, stereoisomeric(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine, and anunidentified impurity. Product: ¹H NMR δ 1.19-1.70 (m, 8H), 1.89-2.05(m, 3H), 2.50 (d, J=6.9 Hz, 2H), 2.56 (m, 1H), 2.78 (m, 2H), 3.44 (m,1H), 7.06 (m, 2H), 7.23 (m, 2H); ¹³C NMR δ 24.39, 25.56, 31.63, 31.76,38.01, 42.61, 49.17, 49.63, 51.74, 62.51, 128.25, 130.42, 131.50,139.16; MS m/z 279.2 (M+1)⁺.

Preparation 7: Preparation of 4-(4-chlorobenzyl)-piperidine

[0182]

Step A: Preparation of 4-(4-chloro-benzylidene)-piperidine-1-carboxylicAcid tert-butyl Ester

[0183]

[0184] The phosphonium salt (10 g) was taken up in THF and placed in anice bath. The KHMDS (42 mL) was added slowly, the ice bath was removed,and the reaction was stirred for 45 minutes at room temperature. Thereaction solution was then cooled to −78° C. and the ketone (4.2 g) wasadded slowly. The reaction was stirred for 30 minutes, the cooling bathwas removed, and the reaction was stirred overnight at room temperature.The reaction solution was poured into a saturated NH₄Cl (100 mL)solution, the layers were separated, the aqueous layer was washed twicewith EtOAc, the organic layers were combined, dried (MgSO₄), andconcentrated to ˜40 mL. The solution was diluted with hexane andfiltered to remove the majority of the Ph₃PO. Chromatography of thecrude product with 20:1-10:1 hexane:EtOAc gave the product as acolorless oil (4.7 g).

Step B: Preparation of 4-(4-chloro-benzyl)-piperidine-1-carboxylic Acidtert-butyl Ester

[0185]

[0186] The protected piperidine (10 g) was dissolved in EtOAc (100 mL),the PtO₂ was added, and the mixture was stirred rapidly under H₂ for 3hours. The mixture was filtered through celite and concentrated. Thecrude product was taken up in hot hexane, filtered and allowed tocrystallize. The product was recrystallized with hot hexane to yield theclean product (8.0 g). Additional product was isolated form the motherliquor.

Step C: Preparation of 4-(4-chlorobenzyl)piperidine

[0187]

[0188] Methanol (400 mL) was placed in an ice bath and AcCl (60 mL) wasadded. After the addition was completed the solution was stirred at roomtemperature for one hour. The protected piperidine (62.8 g) was addedand the solution was stirred at room temperature overnight. The reactionsolution was concentrated to ˜70 mL (when product first started toprecipitate out), diluted with ether (500 mL), and the product wascollected by filtration (44.9 g). An additional 3.1 g of the product wascollected from the mother liquor.

Example 1

[0189] The following compound was prepared using General Procedure E,with the appropriate amine Ia and carboxylic acid.

[0190] Cyclohexanecarboxylic acid{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide

Example 2

[0191] The following compounds were prepared using General Procedure K,with the appropriate amine Ia and isocyanate R⁴N═C═O.

[0192]1-Allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

[0193]1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;

[0194]1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;and

[0195]3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)propionicacid ethyl ester.

Example 3

[0196] The following compounds were prepared using General Procedure J,with the appropriate amine Ia and isocyanate R⁴N═C═O.

[0197]{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea.

Example 4 Formulation Examples

[0198] The following are representative pharmaceutical Formulationscontaining a compound of Formula (I).

[0199] Tablet Formulation

[0200] The following ingredients are mixed intimately and pressed intosingle scored tablets. Quantity per Ingredient tablet, mg compound ofthis invention 400 cornstarch 50 croscarmellose sodium 25 lactose 120magnesium stearate 5

[0201] Capsule Formulation

[0202] The following ingredients are mixed intimately and loaded into ahard-shell gelatin capsule. Quantity per Ingredient capsule, mg compoundof this invention 200 lactose, spray-dried 148 magnesium stearate 2

[0203] Suspension Formulation

[0204] The following ingredients are mixed to form a suspension for oraladministration. Ingredient Amount compound of this invention 1.0 gfumaric acid 0.5 g sodium chloride 2.0 g methyl paraben 0.15 g propylparaben 0.05 g granulated sugar 25.5 g sorbit (70% solution) 12.85 gVeegum K (Vanderbilt Co.) 1.0 g flavoring 0.035 ml colorings 0.5 mgdistilled water q.s. to 100 ml

[0205] Injectable Formulation

[0206] The following ingredients are mixed to form an injectableFormulation. Ingredient Amount compound of this invention 0.2 g sodiumacetate buffer solution, 0.4 M 2.0 ml HCl (1N) or NaOH (1N) q.s. tosuitable pH water (distilled, sterile) q.s. to 20 ml

[0207] Liposomal Formulation

[0208] The following ingredients are mixed to form a liposomalFormulation. Ingredient Amount compound of this invention 10 mgL-.alpha.-phosphatidylcholine 150 mg tert-butanol 4 ml

[0209] Freeze dry the sample and lyophilize overnight. Reconstitute thesample with 1 ml 0.9% saline solution. Liposome size can be reduced bysonication.

Example 5 CCR-3 Receptor Binding Assay—In Vitro

[0210] The CCR-3 antagonistic activity of the compounds of the inventionwas determined by their ability to inhibit the binding of ¹²⁵ I eotaxinto CCR-3 L1.2 transfectant cells (see Ponath, P. D. et al., J. Exp.Med., Vol. 183, 2437-2448, (1996)).

[0211] The assay was performed in Costar 96-well polypropylene roundbottom plates. Test compounds were dissolved in DMSO and then dilutedwith binding buffer (50 mM HEPES, 1 mM CaCl.sub.2, 5 mM MgCl₂, 0.5%bovine serum albumin (BSA), 0.02% sodium azide, pH 7.24) such that thefinal DMSO concentration was 2%. 25 μl of the test solution or onlybuffer with DMSO (control samples) was added to each well, followed bythe addition of 25 μl of ¹²⁵I-eotaxin (100 pmol) (NEX314, New EnglandNuclear, Boston, Mass.) and 1.5×10⁵ of the CCR-3 L1.2 transfected cellsin 25 μl binding buffer. The final reaction volume was 75 μl.

[0212] After incubating the reaction mixture for 1 hour at roomtemperature, the reaction was terminated by filtering the reactionmixture through polyethylenimine treated Packard Unifilter GF/C filterplate (Packard, Chicago, Ill.). The filters were washed four times withice cold wash buffer containing 10 mm HEPES and 0.5M sodium chloride (pH7.2) and dried at 65° C. for approximately 10 minutes. 25 μl/well ofMicroscint-20® scintillation fluid (Packard) was added and theradioactivity retained on the filters was determined by using thePackard TopCount®.

[0213] Compounds of this invention were active in this assay. CompoundIC50 (μM)

2.9116

0.3975

0.4993

0.8846

0.5071

0.7289

Example 6 Inhibition of Eotaxin Mediated Chemotaxis of CCR-3 L1.2Transfectant Cells—In Vitro Assay

[0214] The CCR-3 antagonistic activity of the compounds of thisinvention can be determined by measuring the inhibition of eotaxinmediated chemotaxis of the CCR-3 L1.2 transfectant cells, using a slightmodification of the method described in Ponath, P. D. et al., J. Clin.Invest. 97: 604-612 (1996). The assay is performed in a 24-wellchemotaxis plate (Costar Corp., Cambridge, Mass.). CCR-3 L1.2transfectant cells are grown in culture medium containing RPMI 1640, 10%Hyclone® fetal calf serum, 55 mM 2-mercaptoethanol and Geneticin 418(0.8 mg/ml). 18-24 hours before the assay, the transfected cells aretreated with n-butyric acid at a final concentration of 5 mM/1×10⁶cells/ml, isolated and resuspended at 1×10⁷ cells/ml in assay mediumcontaining equal parts of RPMI 1640 and Medium 199 (M 199) with 0.5%bovine serum albumin.

[0215] Human eotaxin suspended in phosphate buffered saline at 1 mg/mlis added to bottom chamber in a final concentration of 100 nm. Transwellculture inserts (Costar Corp., Cambridge, Mass.) having 3 micron poresize are inserted into each well and L1.2 cells (1×10⁶) are added to thetop chamber in a final volume of 100 μl. Test compounds in DMSO areadded both to the top and bottom chambers such that the final DMSOvolume is 0.5%. The assay is performed against two sets of controls. Thepositive control contained cells with no test compound in the topchamber and only eotaxin in the lower chamber. The negative controlcontains cells with no test compound in the top chamber and neithereotaxin nor test compound in lower chamber. The plate is incubated at37° C. After 4 hours, the inserts are removed from the chambers and thecells that have migrated to the bottom chamber are counted by pipettingout 500 μl of the cell suspension from the lower chamber to 1.2 mlCluster tubes (Costar) and counting them on a FACS for 30 seconds.

Example 7 Inhibition of Eotaxin Mediated Chemotaxis of HumanEosinophils—In Vitro Assay

[0216] The ability of compounds of the invention to inhibit eotaxinmediated chemotaxis of human eosinophils can be assessed using a slightmodification of procedure described in Carr, M. W. et al., Proc. Natl.Acad. Sci. USA, 91: 3652-3656 (1994). Experiments are performed using 24well chemotaxis plates (Costar Corp., Cambridge, Mass.). Eosinophils areisolated from blood using the procedure described in PCT Application,Publication No. WO 96/22371. The endothelial cells used are theendothelial cell line ECV 304 obtained from European Collection ofAnimal Cell Cultures (Porton Down, Salisbury, U.K.). Endothelial cellsare cultured on 6.5 mm diameter Biocoat.RTM. Transwell tissue cultureinserts (Costar Corp., Cambridge, Mass.) with a 3.0 μM pore size.Culture media for ECV 304 cells consists of M199, 10% Fetal Calf Serum,L-glutamine and antibiotics. Assay media consists of equal parts RPMI1640 and M199, with 0.5% BSA. 24 hours before the assay 2×10⁵ ECV 304cells are plated on each insert of the 24-well chemotaxis plate andincubated at 37° C. 20 nM of eotaxin diluted in assay medium is added tothe bottom chamber. The final volume in bottom chamber is 600 μl. Theendothelial coated tissue culture inserts are inserted into each well.10⁶ eosinophil cells suspended in 100 μl assay buffer are added to thetop chamber. Test compounds dissolved in DMSO are added to both top andbottom chambers such that the final DMSO volume in each well was 0.5%.The assay is performed against two sets of controls. The positivecontrol contains cells in the top chamber and eotaxin in the lowerchamber. The negative control contains cells in the top chamber and onlyassay buffer in the lower chamber. The plates are incubated at 37° C. in5% CO₂/95% air for 1-1.5 hours.

[0217] The cells that migrate to the bottom chamber are counted usingflow cytometry. 500 μl of the cell suspension from the lower chamber areplaced in a tube, and relative cell counts are obtained by acquiringevents for a set time period of 30 seconds.

Example 8 Inhibition of Eosinophil Influx Into the Lungs of OvalbuminSensitized Balb/c Mice by CCR-3 Antagonist—In Vivo Assay

[0218] The ability of the compounds of the invention to inhibitleukocyte infiltration into the lungs can be determined by measuring theinhibition of eosinophil accumulation into the bronchioalveolar lavage(BAL) fluid of Ovalbumin (OA)-sensitized balb/c mice after antigenchallenge by aerosol. Briefly, male balb/c mice weighing 20-25 g aresensitized with OA (10 μg in 0.2 ml aluminum hydroxide solution)intraperitoneally on days 1 and 14. After a week, the mice are dividedinto ten groups. Test compound or only vehicle (control group) oranti-eotaxin antibody (positive control group) is administered eitherintraperitoneally, subcutaneously or orally. After 1 hour, the mice areplaced in a Plexiglass box and exposed to OA aerosol generated by aPARISTAR.TM. nebulizer (PARI, Richmond, Va.) for 20 minutes. Mice whichhave not been sensitized or challenged are included as a negativecontrol. After 24 or 72 hours, the mice are anesthetized (urethane,approx. 1 g/kg, i.p.), a tracheal cannula (PE 60 tubing) is inserted andthe lungs are lavaged four times with 0.3 ml PBS. The BAL fluid istransferred into plastic tubes and kept on ice. Total leukocytes in a 20μl aliquot of the BAL fluid is determined by Coulter Counter.TM.(Coulter, Miami, Fla.). Differential leukocyte counts are made onCytospin.TM. preparations which have been stained with a modifiedWright's stain (DiffQuick.TM.) by light microscopy using standardmorphological criteria.

[0219] The foregoing invention has been described in some detail by wayof illustration and example, for purposes of clarity and understanding.It will be obvious to one of skill in the art that changes andmodifications may be practiced within the scope of the appended claims.Therefore, it is to be understood that the above description is intendedto be illustrative and not restrictive. The scope of the inventionshould, therefore, be determined not with reference to the abovedescription, but should instead be determined with reference to thefollowing appended claims, along with the full scope of equivalents towhich such claims are entitled.

[0220] All patents, patent applications and publications cited in thisapplication are hereby incorporated by reference in their entirety forall purposes to the same extent as if each individual patent, patentapplication or publication were so individually denoted.

We claim:
 1. A compound of Formula (I):

wherein: R¹ is (C₁-C₂)alkylene; R² is optionally substituted phenyl; R³is hydrogen, alkyl, acyl, aryl, or arylalkyl; ring A is a cycloalkyl,heterocyclyl, or optionally substituted phenyl; D is N or C—R^(b); L is—C(═O)—, —C(═S)—, —SO₂—, —C(═O)N(R_(a))—, —C(═S) N(R_(a))—, —SO₂N(R_(a))—, —C(═O)O—, —C(═S)O—, —S(═O)₂O—; R⁴ is alkyl, cycloalkyl,alkenyl, alkynyl, heteroalkyl or acylalkyl; R_(a) is hydrogen, alkyl,acyl, aryl, arylalkyl, alkoxycarbonyl, or benzyloxycarbonyl; and R^(b)is hydrogen or alkyl; and prodrugs, individual isomers, racemic andnon-racemic mixtures of isomers, and pharmaceutically acceptable saltsand solvates thereof.
 2. The compound of claim 1, which is a compound ofFormula (II):

wherein R¹-R⁴, A, D, and L have any of the values described in claim 1.3. The compound of claim 1, which is a compound of Formula (III):

wherein R¹-R⁴, A, D and L have any of the values described in claim 1.4. The compound of claim 1 wherein R¹ is methylene.
 5. The compound ofclaim 2 where R¹ is methylene.
 6. The compound of claim 3 where R¹ ismethylene.
 7. The compound of claim 1 wherein R² is 4-chlorophenyl or3,4-dichlorophenyl.
 8. The compound of claim 2 wherein R² is4-chlorophenyl or 3,4-dichlorophenyl.
 9. The compound of claim 3 whereinR² is 4-chlorophenyl or 3,4-dichlorophenyl.
 10. The compound of claim 1wherein R³ is hydrogen.
 11. The compound of claim 2 wherein R³ ishydrogen.
 12. The compound of claim 3 wherein R³ is hydrogen.
 13. Thecompound of claim 1 wherein L is —C(═O)—, —SO₂—, —C(═O)N(R_(a))—,—C(═S)N(R_(a))—, or —C(═O)O—.
 14. The compound of claim 2 wherein L is—C(═O)—, —SO₂—, —C(═O)N(R_(a))—, —C(═S)N(R_(a))—, or —C(═O)O—.
 15. Thecompound of claim 3 wherein L is —C(═O)—, —SO₂—, —C(═O)N(R_(a))—,—C(═S)N(R_(a))—, or —C(═O)O—.
 16. The compound of claim 1 wherein L is—C(═O)—.
 17. The compound of claim 2 wherein L is —C(═O)—.
 18. Thecompound of claim 3 wherein L is —C(═O)—.
 19. The compound of claim 1wherein L is —C(═O)N(R_(a))—.
 20. The compound of claim 2 wherein L is—C(═O)N(R_(a))—.
 21. The compound of claim 3 wherein L is—C(═O)N(R_(a))—.
 22. The compound of claim 1 which is a compound offormula (IV):

wherein R³-R⁴, A, D and L have any of the values described in claim 1.23. The compound of claim 1 which is a compound of formula (V):

wherein R⁴ and D have any of the values defined in claim
 1. 24. Thecompound of claim 1 which is a compound of formula (VI):

wherein R⁴ and D have any of the values defined in claim
 1. 25. Thecompound of claim 1 which is a compound of formula (VII):

wherein R⁴ and D have any of the values defined in claim
 1. 26. Thecompound of claim 1 which is a compound of formula (VIII):

wherein R⁴ and D have any of the values defined in claim 1; and R^(x) ishydrogen, alkyl, acyl, alkylsulfonyl, aminosulfonyl,(alkylamino)sulfonyl, (dialkylamino)sulfonyl, carbamoyl,(alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl)alkyl,(alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl.
 27. Thecompound of claim 1 which is a compound of formula (IX):

wherein R⁴ and D have any of the values defined in claim
 1. 28. Thecompound of claim 1 which is a compound of formula (X):

wherein R⁴ and D have any of the values defined in claim
 1. 29. Thecompound of claim 1 wherein R⁴ is cyclohexyl, allyl, isopropyl, n-butyl,or 2-(ethoxycarbonyl)ethyl.
 30. The compound of claim 1 wherein A is acyclopentyl.
 31. The compound, Cyclohexanecarboxylic acid{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide;{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea;1-Allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)propionicacid ethyl ester; or a salt thereof.
 32. A composition containing atherapeutically effective amount of a compound of claim 1, or a saltthereof; and an excipient.
 33. A method of treatment of a disease in amammal treatable by administration of a CCR-3 receptor antagonist,comprising administering to the mammal a therapeutically effectiveamount of a compound of Formula (I) as described in claim 1, or a saltthereof.
 34. The method of claim 33 wherein the disease is asthma.
 35. Acompound as described in claim 1, or a salt thereof for use in medicaltherapy or diagnosis.
 36. The use of a compound of Formula (I) asdescribed in claim 1, or a salt thereof; for the manufacture of amedicament useful for treating a disease in a mammal treatable byadministration of a CCR-3 receptor antagonist.